LOCUS       Exported                7391 bp ds-DNA     circular SYN 06-DEC-2020
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7391)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Jan 12, 2023 from SnapGene 4.3.6
            https://www.snapgene.com
COMMENT     Insert from 80-pCR8-3xFLAG_3xNLS_dPspCas13b(AAAA)_2xNLS-Clover 652 
            to 5114
            Backbone from 
            pAC90-pmaxGW-D55-60,22-61-69,23-91-7,28-135-11,29-67-30,30-105-27 
            2650 to 967
FEATURES             Location/Qualifiers
     source          1..7391
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1..947
                     /organism="pmaxCloning"
                     /mol_type="other DNA"
                     /label=source:pmaxCloning
     source          960..967
                     /organism="pLenti6/Ubc/V5-DEST Gateway (ViraPower)"
                     /mol_type="other DNA"
                     /label=source:pLenti6/Ubc/V5-DEST Gateway (ViraPower)
     source          join(968..991,5420..5430)
                     /organism="pCR8/GW/TOPO"
                     /mol_type="other DNA"
                     /label=source:pCR8/GW/TOPO
     source          4644..5396
                     /organism="pcDNA3-Clover"
                     /mol_type="other DNA"
                     /label=source:pcDNA3-Clover
     source          5431..5551
                     /organism="pLenti6/Ubc/V5-DEST Gateway (ViraPower)"
                     /mol_type="other DNA"
                     /label=source:pLenti6/Ubc/V5-DEST Gateway (ViraPower)
                     /label=source:pLenti6/Ubc/V5-DEST Gateway (ViraPower)(1)
     source          5564..7391
                     /organism="pmaxCloning"
                     /mol_type="other DNA"
                     /label=source:pmaxCloning
                     /label=source:pmaxCloning(1)
     promoter        163..715
                     /label=CMV_immearly_promoter
     misc_feature    218..505
                     /label=CAG_enhancer
     promoter        661..742
                     /label=CMV_promoter
     misc_feature    672..692
                     /label=CMV_fwd_primer
     misc_feature    813..945
                     /label=chim_int
     misc_feature    960..967
                     /label=attR1
     misc_feature    complement(968..984)
                     /label=attL1
     misc_feature    987..991
                     /label=EcoRIHS
     misc_feature    1008..1076
                     /label=3xFLAG
     misc_feature    join(1098..1169,1176..1187)
                     /label=3xNLS
     CDS             1248..2334
                     /codon_start=1
                     /label=dPspCas13b
                     /translation="MNIPALVENQKKYFGTYSVMAMLNAQTVLDHIQKVADIEGEQNEN
                     NENLWFHPVMSHLYNAKNGYDKQPEKTMFIIERLQSYFPFLKIMAENQREYSNGKYKQN
                     RVEVNSNDIFEVLKRAFGVLKMYRDLTNAYKTYEEKLNDGCEFLTSTEQPLSGMINNYY
                     TVALRNMNERYGYKTEDLAFIQDKRFKFVKDAYGKKKSQVNTGFFLSLQDYNGDTQKKL
                     HLSGVGIALLICLFLDKQYINIFLSRLPIFSSYNAQSEERRIIIRSFGINSIKLPKDRI
                     HSEKSNKSVAMDMLNEVKRCPDELFTTLSAEKQSRFRIISDDHNEVLMKRSSDRFVPLL
                     LQYIDYGKLFDHIRFHVNMGKL"
     misc_feature    2367..4517
                     /label=dPspCas13b
                     /label=dPspCas13b(1)
     misc_feature    4542..4589
                     /label=2xNLS
     misc_feature    4644..5396
                     /label=mClover
     misc_feature    5421..5430
                     /label=attL2
     misc_feature    5498..5539
                     /label=V5
     misc_feature    complement(5498..5517)
                     /label=AC_V5_R
     misc_feature    complement(5565..5584)
                     /label=EBV_rev_primer
     rep_origin      complement(5858..6476)
                     /direction=LEFT
                     /label=pBR322_origin
     CDS             complement(6541..7386)
                     /codon_start=1
                     /label=ORF frame 3
                     /translation="MGGDRDETNPDKCFNNIMIEQDGLHAGSPAAWVERLFGYDWAQQT
                     IGCSDAAVFRLSAQGRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEA
                     GRDWLLLGEVPGQDLLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERART
                     RMEAGLVDQDDLDEEHQGLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSG
                     FIDCGRLGVADRYQDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF
                     "
     gene            complement(6544..7323)
                     /gene="NeoR/KanR (variant)"
                     /label=NeoR/KanR (variant)
                     /note="/translation=MGGDRDETNPDKCFNNIMIEQDGLHAGSPAAWVERLFGY
                      DWAQQ 
                     TIGCSDAAVFRLSAQGRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVT 
                     EAGRDWLLLGEVPGQDLLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIER 
                     ARTRMEAGLVDQDDLDEEHQGLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENG 
                     RFSGFIDCGRLGVADRYQDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRL 
                     LDEFF*"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagg
      721 tcgtttagtg aaccgtcaga tcactagtag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctc gactgatcac aggtaagtat caaggttaca agacaggttt
      841 aaggaggcca atagaaactg ggcttgtcga gacagagaag attcttgcgt ttctgatagg
      901 cacctattgg tcttactgac atccactttg cctttctctc cacagggaaa aaacaattga
      961 caagtttgta caaaaaagca ggctccgaat tcaccggtgc cgccaccatg gactacaagg
     1021 atcacgacgg tgactataag gatcatgaca tcgactataa ggacgatgac gataagatcg
     1081 atggcggagg cggatctgat ccaaaaaaga agagaaaggt agatccaaaa aagaagagaa
     1141 aggtagatcc aaaaaagaag agaaaggtag gatctaccgg atctagaaac gatggtggtg
     1201 gtggaagcgg gggtgggggc agcggtggag ggggaagcgg gcgcgccatg aacatccccg
     1261 ctctggtgga aaaccagaag aagtactttg gcacctacag cgtgatggcc atgctgaacg
     1321 ctcagaccgt gctggaccac atccagaagg tggccgatat tgagggcgag cagaacgaga
     1381 acaacgagaa tctgtggttt caccccgtga tgagccacct gtacaacgcc aagaacggct
     1441 acgacaagca gcccgagaaa accatgttca tcatcgagcg gctgcagagc tacttcccat
     1501 tcctgaagat catggccgag aaccagagag agtacagcaa cggcaagtac aagcagaacc
     1561 gcgtggaagt gaacagcaac gacatcttcg aggtgctgaa gcgcgccttc ggcgtgctga
     1621 agatgtacag ggacctgacc aacgcataca agacctacga ggaaaagctg aacgacggct
     1681 gcgagttcct gaccagcaca gagcaacctc tgagcggcat gatcaacaac tactacacag
     1741 tggccctgcg gaacatgaac gagagatacg gctacaagac agaggacctg gccttcatcc
     1801 aggacaagcg gttcaagttc gtgaaggacg cctacggcaa gaaaaagtcc caagtgaata
     1861 ccggattctt cctgagcctg caggactaca acggcgacac acagaagaag ctgcacctga
     1921 gcggagtggg aatcgccctg ctgatctgcc tgttcctgga caagcagtac atcaacatct
     1981 ttctgagcag gctgcccatc ttctccagct acaatgccca gagcgaggaa cggcggatca
     2041 tcatcagatc cttcggcatc aacagcatca agctgcccaa ggaccggatc cacagcgaga
     2101 agtccaacaa gagcgtggcc atggatatgc tcaacgaagt gaagcggtgc cccgacgagc
     2161 tgttcacaac actgtctgcc gagaagcagt cccggttcag aatcatcagc gacgaccaca
     2221 atgaagtgct gatgaagcgg agcagcgaca gattcgtgcc tctgctgctg cagtatatcg
     2281 attacggcaa gctgttcgac cacatcaggt tccacgtgaa catgggcaag ctgagatacc
     2341 tgctggcggc cgccgctacc tgcatcgacg gccagaccag agtcagagtg atcgagcagc
     2401 ccctgaacgg cttcggcaga ctggaagagg ccgagacaat gcggaagcaa gagaacggca
     2461 ccttcggcaa cagcggcatc cggatcagag acttcgagaa catgaagcgg gacgacgcca
     2521 atcctgccaa ctatccctac atcgtggaca cctacacaca ctacatcctg gaaaacaaca
     2581 aggtcgagat gtttatcaac gacaaagagg acagcgcccc actgctgccc gtgatcgagg
     2641 atgatagata cgtggtcaag acaatcccca gctgccggat gagcaccctg gaaattccag
     2701 ccatggcctt ccacatgttt ctgttcggca gcaagaaaac cgagaagctg atcgtggacg
     2761 tgcacaaccg gtacaagaga ctgttccagg ccatgcagaa agaagaagtg accgccgaga
     2821 atatcgccag cttcggaatc gccgagagcg acctgcctca gaagatcctg gatctgatca
     2881 gcggcaatgc ccacggcaag gatgtggacg ccttcatcag actgaccgtg gacgacatgc
     2941 tgaccgacac cgagcggaga atcaagagat tcaaggacga ccggaagtcc attcggagcg
     3001 ccgacaacaa gatgggaaag agaggcttca agcagatctc cacaggcaag ctggccgact
     3061 tcctggccaa ggacatcgtg ctgtttcagc ccagcgtgaa cgatggcgag aacaagatca
     3121 ccggcctgaa ctaccggatc atgcagagcg ccattgccgt gtacgatagc ggcgacgatt
     3181 acgaggccaa gcagcagttc aagctgatgt tcgagaaggc ccggctgatc ggcaagggca
     3241 caacagagcc tcatccattt ctgtacaagg tgttcgcccg cagcatcccc gccaatgccg
     3301 tcgagttcta cgagcgctac ctgatcgagc ggaagttcta cctgaccggc ctgtccaacg
     3361 agatcaagaa aggcaacaga gtggatgtgc ccttcatccg gcgggaccag aacaagtgga
     3421 aaacacccgc catgaagacc ctgggcagaa tctacagcga ggatctgccc gtggaactgc
     3481 ccagacagat gttcgacaat gagatcaagt cccacctgaa gtccctgcca cagatggaag
     3541 gcatcgactt caacaatgcc aacgtgacct atctgatcgc cgagtacatg aagagagtgc
     3601 tggacgacga cttccagacc ttctaccagt ggaaccgcaa ctaccggtac atggacatgc
     3661 ttaagggcga gtacgacaga aagggctccc tgcagcactg cttcaccagc gtggaagaga
     3721 gagaaggcct ctggaaagag cgggcctcca gaacagagcg gtacagaaag caggccagca
     3781 acaagatccg cagcaaccgg cagatgagaa acgccagcag cgaagagatc gagacaatcc
     3841 tggataagcg gctgagcaac agccggaacg agtaccagaa aagcgagaaa gtgatccggc
     3901 gctacagagt gcaggatgcc ctgctgtttc tgctggccaa aaagaccctg accgaactgg
     3961 ccgatttcga cggcgagagg ttcaaactga aagaaatcat gcccgacgcc gagaagggaa
     4021 tcctgagcga gatcatgccc atgagcttca ccttcgagaa aggcggcaag aagtacacca
     4081 tcaccagcga gggcatgaag ctgaagaact acggcgactt ctttgtgctg gctagcgaca
     4141 agaggatcgg caacctgctg gaactcgtgg gcagcgacat cgtgtccaaa gaggatatca
     4201 tggaagagtt caacaaatac gaccagtgca ggcccgagat cagctccatc gtgttcaacc
     4261 tggaaaagtg ggccttcgac acataccccg agctgtctgc cagagtggac cgggaagaga
     4321 aggtggactt caagagcatc ctgaaaatcc tgctgaacaa caagaacatc aacaaagagc
     4381 agagcgacat cctgcggaag atccggaacg ccttcgatgc aaacaattac cccgacaaag
     4441 gcgtggtgga aatcaaggcc ctgcctgaga tcgccatgag catcaagaag gcctttgggg
     4501 agtacgccat catgaagggc ggccgcggag ggggtggaag tgacccaaag aagaagcgca
     4561 aagtggatcc taaaaagaaa agaaaggtag ggggaggcgg ttccggtggc ggcggaagcg
     4621 gaggtggagg atcagggccg gccatggtga gcaagggcga ggagctgttc accggggtgg
     4681 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtccgcggcg
     4741 agggcgaggg cgatgccacc aacggcaagc tgaccctgaa gttcatctgc accaccggca
     4801 agctgcccgt gccctggccc accctcgtga ccaccttcgg ctacggcgtg gcctgcttca
     4861 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     4921 acgtccagga gcgcaccatc tctttcaagg acgacggtac ctacaagacc cgcgccgagg
     4981 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     5041 aggacggcaa catcctgggg cacaagctgg agtacaactt caacagccac aacgtctata
     5101 tcacggccga caagcagaag aacggcatca aggctaactt caagatccgc cacaacgttg
     5161 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     5221 ccgtgctgct gcccgacaac cactacctga gccatcagtc cgccctgagc aaagacccca
     5281 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg attacacatg
     5341 gcatggacga gctgtacaag tctagagggc cctattctat agtgtcacct aaatgcatcg
     5401 attgattaat taagaattcg acccagcttt cttgtacaaa gtggttgata tccagcacag
     5461 tggcggccgc tcgagtctag agggcccgcg gttcgaaggt aagcctatcc ctaaccctct
     5521 cctcggtctc gattctacgc gtaccggtta gcaattgttt tttcgatgag tttggacaaa
     5581 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     5641 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     5701 tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat
     5761 gtggtactta agagggggag accaaagggc gagacgttaa ggcctcacgt gacatgtgag
     5821 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     5881 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     5941 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     6001 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     6061 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     6121 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     6181 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     6241 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     6301 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     6361 aaagagttgg tagctcttga tccggcaaac aaaccacgct ggtagcggtg gtttttttgt
     6421 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6481 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgccgtc
     6541 tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat
     6601 accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag caatatcacg
     6661 ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac agtcgatgaa
     6721 tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc catgggtcac
     6781 gacgagatcc tcgccgtcgg gcatgctcgc cttgagcctg gcgaacagtt cggctggcgc
     6841 gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt ccatccgagt
     6901 acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag
     6961 cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag gagcaaggtg
     7021 agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc ttcccgcttc
     7081 agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg
     7141 cgctgcctcg tcttgcagtt cattcagggc accggacagg tcggtcttga caaaaagaac
     7201 cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga ttgtctgttg
     7261 tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc
     7321 atcttgttca atcataatat tattgaagca tttatcaggg ttcgtctcgt cccggtctcc
     7381 tcccatgcat g
//
